中文摘要：本文开发了一个新的基于环介导等温扩增（LAMP）技术的转基因玉米和大豆事件的筛选方法。LAMP方法以七个序列为靶标：花椰菜花叶病毒35S启动子、根癌农杆菌CP4（CP4EPSPS）的5-烯醇丙酮酸莽草酸-3-磷酸合成酶基因、草丁膦 N -乙酰转移酶（PAT）、甘露糖-6-磷酸异构酶基因、豌豆核糖1，5-二磷酸羧化酶终止子、Cyr1Ab和Cy1Ac基因的共同序列、GA21结构特异序列。我们为每个靶标设计了新的特定引物集，并使用授权转基因玉米和大豆事件的检测限（LOD）进行评估。LODs的各项指标均<< 0.5%。为了使DNA提取过程简单快速，我们还利用粗细胞裂解液开发了直接LAMP检测方案。整个过程，包括预处理和检测，可以在1小时内完成。
外文摘要：We developed new loop-mediated isothermal amplification (LAMP)-based detection methods for the screening of genetically modified (GM) maize and soybean events. The LAMP methods developed targeted seven sequences: cauliflower mosaic virus 35S promoter; 5-enolpyruvylshikimate-3-phosphate synthase gene from Agrobacterium tumefaciens strain CP4 (cp4epsps); phosphinothricin acetyltransferase (pat) gene; mannose-6-phosphate isomerase gene; Pisum sativum ribulose 1, 5-bisphosphate carboxylase terminator; a common sequence between Cry1Ab and Cry1Ac genes; and a GA21 construct-specific sequence. We designed new specific primer sets for each target, and the limit of detection (LOD) was evaluated using authorized GM maize and soybean events. LODs for each target were <= 0.5%. To make the DNA extraction process simple and rapid, we also developed a direct LAMP detection scheme using crude cell lysates. The entire process, including pretreatments and detection, could be completed within 1 h.
外文关键词：Loop-mediated isothermal amplification (LAMP), Genetically modified (GM), Direct LAMP, Rapid qualitative analysis
作者：Takabatake, Reona; Kagiya, Yukari; Minegishi, Yasutaka; Yeasmin, Sabina; Futo, Satoshi; Noguchi, Akio; Kondo, Kazunari; Mano, Junichi; Kitta, Kazumi
作者单位：National Agriculture and Food Research Organization
期刊名称：FOOD CHEMISTRY
期刊影响因子：4.529
出版年份：2018
出版刊次：6
点击下载：基于环介导等温扩增技术的转基因作物快速筛选检测方法的建立与评价